PS 14-185 - CANCELLED - Microbe surveillance in the amphibian pet trade: results from a pilot study

Abstract: It is widely acknowledged that regional and global trade of live animals contribute to the spread and emergence of pathogens. Yet beyond small portions of large, complex trade networks (e.g., bullfrogs in U.S. ports, exotic amphibians shipped through Hong Kong markets) little is known about how pathogens are amplified or diminished, or even how common they might be. In part, this is because facilities are reluctant to participate in surveillance that might expose problems with disease or trade secrets. As part of a larger effort to understand the amphibian pet trade in the U.S.A., we sent DNA sampling kits to 14 businesses that volunteered to join an anonymous surveillance program in which participant identities were masked through collaboration with a trusted trade group. Kits contained swabs for sampling terrestrial animals and environmental DNA (eDNA) filters to sample aquatic housings. Participants were also provided detailed written and video instructions. Returned samples were pooled by habitat (e.g., container) and tested for the presence of the emerging amphibian pathogens Batrachochytrium salamandrivorans (Bsal), B. dendrobatidis (Bd), and Ranavirus spp. (Rv) as well as the beneficial microbe, Janthinobacterium lividum (Jliv) using standard quantitative real time PCR (qPCR) assays. Of the 14 businesses that were shipped DNA collection supplies, eight returned samples. Bd was found in two facilities, while Bsal, Rv, and Jliv were not detected. A qPCR assay targeting a highly conserved region of vertebrate DNA (EBF3N) was used to validate successful sample collection across all sites. EBF3N titers were consistently low ( < 100 copies) and seemingly unrelated to Bd detection, suggesting this gene target may ultimately be a poor proxy for sample quality. The results of this pilot study highlight the feasibility of sampling the pet trade more comprehensively, as well as an interest in such surveillance within the industry.